A standardized protocol for genetically stable artificial seed production of Ficus religiosa L. using ISSR fingerprinting

Abstract

Ficus religiosa LQuery. has several ornamental, medicinal, and economical applications. The in vivo propagation of this species has shown various limitations. Due to this reason, the present study efforts on genetically uniform artificial seed production from in vitro developed shoot tips of this species. The in vivo shoot tips were cultivated on Murashige and Skoog (MS) media containing different growth regulators. The maximum shoot response (93.67%) and the longest shoot length (3.85cm) were exhibited with 0.5mg L-1 6-furfuryl-amino purine (Kn), 0.2mg L-1 benzyladenine (BA) and 0.1mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) in combination. A treatment of 3% sodium alginate and 75mM calcium chloride having a polymerization time of 15min was exhibited to be superior for artificial seed production of these in vitro grown shoot tips. Artificial seed-derived micro shoots yielded the highest root response (94.44%) and roots per shoot (4.61) with 0.5mg L-1 indole-3-butyric acid (IBA) and 0.1mg L-1 BA in combination on ½-strength MS media. In comparison to 4°C-kept artificial seeds, 24°C-stored artificial seeds had superior germination potential across all storage times. The soil:organic manure (1:1) generated 90% of plantlet survival after 28days of primary hardening than other mixtures tested. The secondary hardening displayed 92% of plant survival after 60days. The banding patterns of ISSR analysis between the mother plant and hardened plants were discovered to be monomorphic. This methodology provides a promising and affordable approach to the large-scale plant productionof this significant species.